Clinical profile of a new monoclonal antibody-based immunoassay for tissue polypeptide antigen

Our preliminary evaluation of a new monoclonal antibody-based assay for tissue polypeptide antigen (TPA) has shown it to be clinically equivalent to the polyclonal antibody-based assay for TPA. The new assay (TPA-M) employs three monoclonal antibodies to epitopes on cytokeratins 8, 18 and 19. This multicenter, multinational study included 266 patients with newly diagnosed carcinomas of the lung, breast, large bowel and urinary bladder. TPA values from the two assays were compared with three other cytokeratin markers (TPS, CYFRA 21-1 and TPACyk) and with the established reference markers for these malignancies (CEA and NSE for lung, CA 15-3 for breast, CEA and CA 19-9 for colorectal tumors). Analysis of receiver operating characteristic (ROC) curves in lung, colorectal and bladder cancer showed similar sensitivities for the two assays, ranging from 50% to 80% with a specificity of 95%. In breast cancer all the markers studied showed poor sensitivity. However, TPA determination by either method could discriminate advanced stage (stages III and IV) from early stage disease (stages 0 to II). TPA showed similar discriminating ability in bladder cancer. On the basis of the results obtained in our patient series, it seems that of the cytokeratin markers studied, TPA and TPA-M are the most sensitive and offer a wide range of clinical applications.     

Mitochondrial complex I defects in aging

According to the 'mitochondrial theory of aging' it is expected that the activity of NADH Coenzyme Q reductase (Complex I) would be most severely affected among mitochondrial enzymes, since mitochondrial DNA encodes for 7 subunits of this enzyme. Being these subunits the site of binding of the acceptor substrate (Coenzyme Q) and of most inhibitors of the enzyme, it is also expected that subtle kinetic changes of quinone affinity and enzyme inhibition could develop in aging before an overall loss of activity would be observed. The overall activity of Complex I was decreased in several tissues from aged rats, nevertheless it was found that direct assay of Complex I using artificial quinone acceptors may underevaluate the enzyme activity. The most acceptable results could be obtained by applying the 'pool equation' to calculate Complex I activity from aerobic NADH oxidation; using this method it was found that the decrease in Complex I activity in mitochondria from old animals was greater than the activity calculated by direct assay of NADH Coenzyme Q reductase. A decrease of NADH oxidation and its rotenone sensitivity was observed in nonsynaptic mitochondria, but not in synaptic 'light' and 'heavy' mitochondria of brain cortex from aged rats. In a study of Complex I activity in human platelet membranes we found that the enzyme activity was unchanged but the titre for half-inhibition by rotenone was significantly increased in aged individuals and proposed this change as a suitable biomarker of aging and age-related diseases.     

Energetics of 3.5 s neural activation in humans: a 31P MR spectroscopy study

No direct information on brain energetics and energy-related compounds in the first seconds of physiological activation has been reported to date. In this study visual cortex high energy phosphate changes were monitored in 11 normal subjects during 3.5 s activation and the following 23.5 s by a simple 31P magnetic resonance spectroscopic method. An intraactivation decrease of phosphocreatine (PCr) was observed in all subjects, with changes in pH in three, one of them also presenting a change in adenosine triphosphate (ATP). In the subgroup of eight subjects without changes in pH, the mean rate of mean PCr decrease (D(PCr)) was 7.24 +/- 0.78%/s, and the postactivation mean rate of mean PCr recovery was <1/2 D(PCr). Short phasic neural activity requires a large amount of energy, i.e., at least three times basal consumption, in agreement with theoretical calculations. Additional energy demands in the visual cortex are several times those measured by positron emission tomography during prolonged stimulation studies, implying that mean energy requirements decrease with increases in duration of stimulation. During short activation, the vascular responses as detected by brain-mapping techniques (BMT) are preceded by an important reduction of the intracellular high-energy phosphate content, which returns to resting values during an interval that corresponds to the poststimulation return of BMT signals to baseline.     

Effects of lipoxygenase and of chemical oxidising agent potassium iodate on rheological properties of durum dough

To study the involvement of lipoxygenase (LOX) and oxidising agent potassium iodate (KIO₃) in rheological behaviour of dough, mixograph and alveograph tests were performed using durum cultivars differing for quality and endogenous levels of LOX. Both tests were done in standard condition and with the adding of KIO₃ and of LOX and/or its substrate, utilised singly or in different combinations. The effects of these treatments were related to change of ‐SH and S–S content in relative mixograph dough. The mixograph and alveograph responses showed that the KIO₃ had an improving effect on dough tenacity and that the adding of LOX system enhanced this trend; on the contrary, the LOX system alone seemed not affect mixing stability. No relation was found between rheological responses and ‐SH and S–S content in the mixograph doughs. The results obtained could suggest that in durum wheat the main effect of LOX could be ascribable to bleaching action because of the co‐oxidation of carotenoid pigments.     

Grinding operation optimization of the CODELCO-Andina concentrator plant

This paper presents results following the application of a sub-optimal control scheme, both through simulation and in situ, from the operation of Section C of the CODELCO-Andina copper concentrator plant. The algorithm permits the determination of the necessary control action at each instant of time in order to maximize a defined plant performance index. The main objective of the algorithm is to maximize the mineral tonnage processed by the section, subject to it not exceeding a predetermined value establishes! for the operation conditions of the mills, while at the same time maintaining constant the mass fraction over 65 mesh (212 [microns]) in the overflow of the hydrocyclones, at a value within the operational requirements of the flotation stage. The performance index is defined in terms of; the percentage of pulp solids fed to the hydrocyclones of each line of ball mills, penalty functions to prevent electric power to the ball mills ,falling below the lower limit (so as not to enter the overload region), the tonnage processed in the section, and, since water is a scarce resource, a term considering the water added to each sump is also included. The scheme is first studied and adjusted in a simulator of a concentrator plant similar to that used in the industrial application. For plant implementation, a PC software program, denoted CONMOL, is developed in TurboPascal for Windows. This software allows plant applications to be carried out through a communications interface. Finally, the results of two tests performed on Section C of the CODELCO-Andina copper concentrator plant are shown where the control is applied over 3 and 5.5 hours respectively.     

The potential transformer bridge with current comparator formeasuring the voltage dependence of compressed-gas capacitors

In this paper, an alternative method for measuring the voltage dependent capacitance variation (VDCV) of compressed-gas capacitors is described. In the proposed method, the VDCV of the capacitor under test is measured by comparing it with a capacitor standard supplied with the secondary (low) voltage of a potential transformer. This transformer has its primary parallel connected with the capacitor under test. The paper presents and criticizes the test results obtained in which an uncertainty of at least 0.5 ppm is achieved     

Not just another self-consistent magnetic recording model

A self-consistent magnetic recording modeling method is presented which has proven useful in recording channel design on thin media. Improvements in the magnetic model and in the mathematical treatment stabilize the iterative process and reduce computer storage requirements. Major and minor media loops are fitted to quickly computable bipolynomials. Mathematical improvements include using a strong band diagonal demagnetization matrix, analytic integration with quadratic magnetization fitting, and Newton-Raphson iteration, which gives rapid convergence without underrelaxation. Quantitative predictions of timing errors in 16-bit modified frequency modulation (MFM) data patterns on a 350-bit/mm, 20-track/mm disk memory are presented, as well as playback amplitudes and saturation currents. Predictions can also be compared with experimental read/write data to determine system parameters. Two examples are given: the inference of the head efficiency and of the effective high-frequency medium squareness.     

Vectors for Cu2+-inducible production of glutathione S-transferase-fusion proteins for single-step purification from yeast

We describe six new yeast episomal vectors which encode glutathione S-transferase (GST) affinity tags. These allow for the production of GST-fusion proteins in Saccharomyces cerevisiae under the control of the CUP1 promoter. Affinity chromatography with glutathione-Sepharose permits convenient purification of the fusion protein from a yeast lysate. The presence of a protease cleavage site facilitates subsequent removal of the GST tag. The expression and single-step purification of both GST and a functional GST-metallothionein fusion from yeast are shown as an example of the application of these vectors.     

The effects of various processing conditions on a protein isolate from Lupinus angustifolius

Lupin protein is a promising ingredient in functional foods because of its purported hypocholesterolaemic and hypotensive activities. In this study a lupin protein isolate from Lupinus angustifolius was thermally and mechanically treated and the effects on its protein profile were determined. As a preliminary step, the main protein components of L. angustifolius were identified, using the canonical proteomic approach, including 2D-separation and mass spectrometry and, whenever necessary, also “de novo peptide sequencing”. Most of the main spots were assigned to the major lupin storage proteins: α-conglutin, β-conglutin, γ-conglutin, and δ-conglutin. The protein degradation induced by the different treatments was studied via differential scanning calorimetry (DSC), 2D-electrophoresis, and mass spectrometry, in order to get the fingerprint of the intact peptides after processing. The results indicate that, even after harsh industrial processing, α-, β- and δ-conglutin are still able to release stable peptides, although they are completely or partially degraded, as shown by the 2D protein profiles and the DSC graphs.     

Effect of ultrasound treatment on properties of gluten-based film

Gluten films obtained in acid conditions display some protein dispersion difficulties. Ultrasound treatment (UT) could represent an interesting strategy for improving gluten film appearance. Different UT exposure times were applied to film-forming dispersion. The aim of this work was to investigate the effect of ultrasound treatment on gluten-based films at molecular and mesoscopic levels. Distribution in gliadin and glutenins was determined with SE-HPLC. The UT improved protein dispersion and final film appearance. Sonication did not lead to large changes in various gluten fractions, which suggests absence of important protein breakdown at the molecular level. Gluten showed high tolerance to UT. Surface properties of untreated and treated films were investigated by contact angle measurement: sonication promoted hydrophilic surface properties.Industrial relevanceBio-based packaging has been receiving increasing attention in view of its beneficial impact on the environment. Among proteins, gluten resulted as a very interesting film-forming material. Gluten films prepared in acid conditions showed problems in protein dispersion. Sonication represents a physical strategy which allowed us to obtain gluten-based films without the addition of chemical additives, such as sodium sulphite.